SynBBB 3D血腦屏障模型芯片，SynBBB Kits and Chips

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SynVivo的SynBBB 3D血腦屏障模型通過模擬與跨血腦屏障（BBB）的內(nèi)皮細(xì)胞通訊的腦組織細(xì)胞的組織切片來重建體內(nèi)微環(huán)境。剪切誘導(dǎo)的內(nèi)皮細(xì)胞緊密連接在Transwell®模型中無法實(shí)現(xiàn)，而在SynBBB模型中使用生理性流體流很容易實(shí)現(xiàn)。緊密變化的形成可以使用SynVivo細(xì)胞阻抗分析儀通過生化或電氣分析（評(píng)估電阻變化）進(jìn)行測量。腦組織細(xì)胞與內(nèi)皮細(xì)胞之間的相互作用在SynBBB分析中很容易觀察到。 Transwell模型不允許實(shí)時(shí)顯示這些細(xì)胞相互作用，這對(duì)于了解BBB微環(huán)境至關(guān)重要。

SynBBB是wei一可以實(shí)現(xiàn)以下功能的體外BBB模型：

準(zhǔn)確的體內(nèi)血液動(dòng)力學(xué)切應(yīng)力
實(shí)時(shí)可視化細(xì)胞和屏障功能
大大減少了成本和時(shí)間
穩(wěn)健易用的協(xié)議

BBB模型的示意圖。頂腔（外通道）用于培養(yǎng)血管（內(nèi)皮細(xì)胞），而基底外側(cè)腔（中央腔）用于培養(yǎng)腦組織細(xì)胞（星形細(xì)胞，周細(xì)胞，神經(jīng)元）。多孔結(jié)構(gòu)使血管細(xì)胞與組織細(xì)胞之間可以進(jìn)行通訊。

SynBBB系統(tǒng)是一個(gè)高度通用的平臺(tái)，可用于調(diào)查：

緊密連接蛋白：確定緊密連接蛋白的水平，即調(diào)節(jié)BBB的小帶閉合蛋白，claudins和occludins。
轉(zhuǎn)運(yùn)蛋白：分析正常和功能異常的血腦屏障中轉(zhuǎn)運(yùn)蛋白的功能（例如Pgp）。
藥物滲透性：評(píng)估治療劑和小分子穿過BBB內(nèi)皮細(xì)胞的實(shí)時(shí)滲透性。
炎癥：了解炎癥反應(yīng)對(duì)血腦屏障調(diào)節(jié)的潛在機(jī)制。
細(xì)胞遷移：可視化并量化免疫細(xì)胞在BBB中的實(shí)時(shí)遷移。
滲透性變化：對(duì)正常和功能異常的血腦屏障進(jìn)行基因組，蛋白質(zhì)組和代謝分析。
神經(jīng)毒性：分析化學(xué)，生物和物理試劑對(duì)血腦屏障細(xì)胞的毒性作用。
神經(jīng)腫瘤學(xué)：研究腫瘤細(xì)胞對(duì)血腦屏障的影響。
根據(jù)您的研究需求，您可以從“基本” SynBBB模型或“ TEER兼容” SynBBB配置中進(jìn)行選擇。

SynBBB基本配置

使用實(shí)時(shí)成像，生化和分子生物學(xué)方法來分析實(shí)驗(yàn)。

具有SynVivo電池阻抗分析儀的TEER兼容SynBBB配置

使用實(shí)時(shí)跨內(nèi)皮電阻（TEER）分析實(shí)驗(yàn)，包括實(shí)時(shí)成像，生化和分子生物學(xué)方法。

SynBBB 3D模型套件組件
可以以試劑盒形式購買運(yùn)行SynBBB分析所需的所有基本組件。根據(jù)個(gè)人研究需求，您可以從SynBBB芯片的“基本”或“ TEER兼容”配置中進(jìn)行選擇。包括所有附件，包括管子，夾子，針頭和注射器。入門工具包還將包括氣動(dòng)啟動(dòng)裝置（運(yùn)行SynBBB分析所需）和細(xì)胞阻抗分析儀（收集SynBBB TEER測量值所需）。

SynVivo’s  SynBBB 3D blood brain barrier model recreates the in vivo microenvironment by emulating a histological slice of brain tissue cells in communication with endothelial cells across the blood brain barrier (BBB). Shear-induced endothelial cell tight junctions, which cannot be achieved in the Transwell® model, are easily achieved in the SynBBB model using physiological fluid flow. Formation of tight changes can be measured using biochemical or electrical analysis (assessing changes in electrical resistance) with the SynVivo Cell Impedance Analyzer. Interactions between brain tissue cells and endothelial cells are readily visualized in the SynBBB assay. Transwell models do not allow real-time visualization of these cellular interactions, which are critical for understanding of the BBB microenvironment.

SynBBB is the only in vitro BBB model that allows:

• Accurate in vivo hemodynamic shear stress
• Real-time visualization of cellular and barrier functionality
• Significant reduction in cost and time
• Robust and easy to use protocols

Schematic of the BBB Model. Apical chamber (outer channels) are for culture of vascular (endothelial cells) while basolateral chamber (central chamber) are for culture of brain tissue cells (astrocytes, pericytes, neurons). Porous architecture enables communication between the vascular and tissue cells.

The SynBBB system is a highly versatile platform for investigation of:

• Tight junction proteins: Determine the levels of tight junction proteins namely zonula occludens, claudins and occludins which regulate the BBB.
• Transporter proteins: Analyze functionality of transporter proteins (e.g. Pgp) in normal and dysfunctional BBB.
• Drug permeability: Evaluate real-time permeability of therapeutics and small molecules across the endothelium of the BBB.
• Inflammation: Understand the underlying mechanisms of  inflammatory responses on the regulation of the BBB.
• Cell migration: Visualize and quantify in real-time migration of immune cells across the BBB.
• Omic changes: Perform genomic, proteomic and metabolic analysis on normal and dysfunctional BBB.
• Neurotoxicity: Analyze toxicity effects of chemical, biological and physical agents on the cells of the BBB.
• Neuro-oncology: Investigate effects of the tumor cells on the BBB.

Depending on your research needs you can select from the “basic” SynBBB model or a “TEER-compatible” SynBBB configuration.

Basic SynBBB Configuration

Analyze experiments using real-time imaging, biochemical and molecular biology methodologies.

TEER Compatible SynBBB Configuration with the SynVivo Cell Impedance Analyzer

Analyze experiments using real-time trans-endothelial electrical resistance (TEER) including real-time imaging, biochemical and molecular biology methodologies.

SynBBB 3D Model Kit Components

All the basic components required to run the SynBBB assay can be purchased in a kit format. Depending on individual research needs you can select from the “basic” or “TEER-compatible” configurations of the SynBBB chip. All accessories including tubing, clamps, needles and syringes are included. Starter kits will also include the pneumatic priming device (required for running SynBBB assays) and the cell impedance analyzer (required for collecting SynBBB TEER measurements).

SynBBB 3D Model Kit Components

All the basic components required to run the SynBBB assay can be purchased in a kit format. Depending on individual research needs you can select from the “basic” or “TEER-compatible” configurations of the SynBBB chip. All accessories including tubing, clamps, needles and syringes are included. Starter kits will also include the pneumatic priming device (required for running SynBBB assays) and the cell impedance analyzer (required for collecting SynBBB TEER measurements).

Kit contents and Description

SynVivo used to create the first neonatal BBB model on a chip

Researchers at Temple University used the SynVivo® SynBBB^TM cell-based in vitro assay platform to model the attributes and functions of the neonatal stage blood-brain barrier (BBB) [1]. The SynBBB model closely mimics the in vivo microenvironment including three-dimensional morphology, cellular interactions and flow characteristics on a microfluidic chip. This work marks the first dynamic in vitro neonatal BBB model that offers real time visualization and analysis and is suitable for studies of BBB function as well as screening of novel therapeutics.

“The work is important because studies of neonatal neuropathologies and development of appropriate therapeutics are hampered by a lack of relevant in vitro models of the neonatal blood-brain barrier,” said Dr. Sudhir Deosarkar, the lead author of this paper.

In the SynBBB assay, which includes a tissue compartment and vascular channels placed side-by-side and separated by an engineered porous barrier, the researchers were able to co-culture neonatal rat brain endothelial cells and rat astrocytes under physiological conditions observed in vivo. The endothelial cells formed a full lumen and exhibited tight junction formation which increased under co-culture with astrocytes. The permeability of small molecules in the developed model was found to in excellent agreement with in vivo observations.

“The real-time visualization capabilities of the SynBBB co-culture platform allowed, for the first time, visualization of astrocyte end-feet and endothelial cell interactions in anin vitro model,” said Prof. Mohammad Kiani who is the senior author of the paper. “This is a unique capability and will help us to understand and develop therapeutics for several developmental disorders and diseases of the brain.”

The PLOS ONE paper shows that in contrast to transwell models, the SynBBB model exhibits significantly improved barrier characteristics similar to in vivo observations.

¹A Novel Dynamic Neonatal Blood-Brain Barrier on a Chip. S. Deosarkar, B. Prabhakarpandian, B. Wang, J.B. Sheffield, B. Krynska, M. Kiani. PLOS ONE, 2015, DOI: 10.1371/journal.pone.0142725

The SynBBB 3D model has been validated in various BBB Assays

Mono-Culture Assays

Shear-induced endothelial cell tight junctions, which cannot be achieved in the Transwell® model, are easily achieved in the SynBBB assay using fluid perfusion. Formation of tight changes can be measured using biochemical or electrical analysis (assessing changes in electrical resistance) with the SynVivo Cell Impedance Analyzer.

Primary endothelial cells are cultured in the vascular channel under physiological fluid flow. Cells are stained for tight junction markers highlighting the increase under fluid flow compared to static conditions. The Cell Impedance Analyzer system is used to measure increases in Ohmic resistance (TEER), associated with the formation of tight junctions.

Top Left Panel: Phase Contrast imaging of brain endothelial cells cultured in the SynBBB model. Bottom Left Panel: Calcein AM and Ethidium homodimer-1 labeled brain endothelial cells indicating a highly viable population of cells in the SynBBB model. Right Panel: Plot highlighting the importance of flow on brain endothelial cells with increased TEER.

Co-Culture with Tissue Cells

Interactions between brain tissue cells and endothelial cells are readily visualized in the SynBBB assay. Transwell models do not allow real-time visualization of these cellular interactions, which are critical for understanding of the physiological environment.

Endothelial cells are cultured under flow in the vascular channel, and the tissue chamber is cultured with primary brain cells, such as astrocytes. Increases in Ohmic resistance across the barrier, measured with the Cell Impedance Analyzer, are associated with tight junction formation across the BBB. Endothelial cells co-cultured with astrocytes form significantly tighter cell junctions compared to mono-cultured endothelial cells.

Left Panel: CD-31 (green) stained endothelial cells and GFAP (red) stained astrocytes. All nucleus are stained with DAPI (blue). Right Panel: Plot highlighting increased TEER with co-culture of endothelial cells and astrocytes.

Real-Time Permeability Assays

Unlike BBB models which are arranged in top to bottom architecture (i.e., Transwell), small molecule transport can be assessed and quantified in real-time across the SynBBB system due to its side-by-side architecture.

A fluorescently-labeled drug molecule of interest is perfused through the vascular channels at physiological  flow rate. Real-time videos are acquired and analyzed to calculate the rate of permeability into the tissue chamber. Different rates of permeability is observed across the BBB due to tight junctions of endothelial cells.

Time-lapse imaging of permeability of small molecules across a tightly formed BBB.

Time-lapse imaging of permeability of small molecules across a leaky BBB.

Real-Time Tight Junction Modulation

SynBBB can be used to model inflammation responses. A pro-inflammatory compound, such as TNF-α, is added to mono-cultured endothelial cells to modulate the tight junctions, followed by a period of recovery under perfusion flow. Electrical resistance measurements provide a non-invasive method for real-time monitoring of tight junctions.

Modulation of Inflammation responses in SynBBB model. TNF-alpha induced leakiness in the BBB measured by changes in the resistance across the endothelial cells. Removal of TNF-alpha followed by media perfusion under physiological flow conditions enables recovery of the tight junction leading to increased tight junction formation. Static cells maintain a constant resistance due to lack of tight junctions.

Includes consumables (10 chips, 100ft tubing, 25 slide clamps, 50 blunt tip needles and 50 1 ml syringes). Starter kits will also include the pneumatic priming device (required for seeding cells) and the cell impedance analyzer (for TEER configuration only).

Cat#

Product Name

Literature

Price

402002

SynBBB 3D Model Starter Kit (basic configuration)

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402004

SynBBB 3D Model Starter Kit (TEER configuration)

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Note: Does not include other required consumables such as cells, media and matrix.
Laboratory equipment required includes incubators, inverted microscopes, and syringe pumps.

SynBBB 3D Model – Assay Kits

Includes consumables (10 chips, 100ft tubing, 25 slide clamps, 50 blunt tip needles and 50 1 ml syringes).

Cat#

Product Name

Literature

402001

SynBBB 3D Model Assay Kit (basic configuration)

view

402003

SynBBB 3D Model Assay Kit (TEER configuration)

view

Note: Does not include other required consumables such as cells, media and matrix.
Laboratory equipment required includes incubators, inverted microscopes, and syringe pumps.

SynBBB 3D Model – Chips

Purchase single chips.

Cat#

Product Name

Literature

Price

102005-SB

SynBBB 3D Model Chip (basic configuration)

view

102015-SB

SynBBB 3D Model Chip (TEER configuration)

view

Note: Does not include other required consumables such as tubing, clamps, syringes, needles, cells, media and matrix.
Laboratory equipment required includes incubators, inverted microscopes, and syringe pumps.

Includes consumables (10 chips, 100ft tubing, 25 slide clamps, 50 blunt tip needles and 50 1 ml syringes). Starter kits will also include the pneumatic priming device (required for seeding cells) and the cell impedance analyzer (for TEER configuration only).

Cat#

Product Name

Literature

Price

402002

SynBBB 3D Model Starter Kit (basic configuration)

view

$1,323ADD TO CART

402004

SynBBB 3D Model Starter Kit (TEER configuration)

view

$2,583ADD TO CART

Note: Does not include other required consumables such as cells, media and matrix.
Laboratory equipment required includes incubators, inverted microscopes, and syringe pumps.

SynBBB 3D Model – Assay Kits

Includes consumables (10 chips, 100ft tubing, 25 slide clamps, 50 blunt tip needles and 50 1 ml syringes).

Cat#

Product Name

Literature

Price

402001

SynBBB 3D Model Assay Kit (basic configuration)

view

$1,040ADD TO CART

402003

SynBBB 3D Model Assay Kit (TEER configuration)

view

$1,354ADD TO CART

Note: Does not include other required consumables such as cells, media and matrix.
Laboratory equipment required includes incubators, inverted microscopes, and syringe pumps.

SynBBB 3D Model – Chips

Purchase single chips.

Cat#

Product Name

Literature

Price

102005-SB

SynBBB 3D Model Chip (basic configuration)

view

$103ADD TO CART

102015-SB

SynBBB 3D Model Chip (TEER configuration)

view

$135ADD TO CART

Note: Does not include other required consumables such as tubing, clamps, syringes, needles, cells, media and matrix.
Laboratory equipment required includes incubators, inverted microscopes, and syringe pumps.